MARA University of Technology

Faculty of Applied Science

Bachelor of Science Applied Chemistry

CHM 510

Experiment 7

ANALYSIS OF CHLORPYRIFOS IN WATER BY SOLID-PHASE

EXTRACTION(SPE) AND GAS CHROMATOGRAPHY- ELECTRON CAPTURE
DETECTOR(GC-ECD)

Group members:

Muhammad Azri Azizi Bin Zakaria 2018272708

Nur Fatin Najihah Binti Ismail 2018438612

Nur Izzatie Binti Halil 2018249186

Syamimi Yasmin Binti Md Raily 2018272956

Prepared for:

Dr Intan Soraya

Date of experiment:

08 October 2019

Date of submission:

15 October 2019
 INDEX FULL MARKS MARKS
Objective 1.0

Introduction 2.0

Procedure 1.0

Result and Observations 3.0

Calculations 3.0

Discussion 2.0

Conclusion 1.0

Questions 3.0

References 1.0

Jotter 2.0

Lab techniques 1.0

OBJECTIVE:

To analyse compound in the waste water using Solid-Phase Extraction (SPE) and Gas
Chromatography-Electron Capture Detector (ECD).

INTRODUCTION:

Figure 1: chlorpyrifos

Chlorpyrifos is a crystalline organophosphate insecticide that acts on the nervous system of

insects by inhibiting acetylcholinesterase. Chlorpyrifos is moderately toxic to humans and the
exposure will cause neurological effects such as persistent developmental disorders
autoimmune disorders and children mental development retard due to the exposure during
pregnancy. In agriculture, chlorpyrifos remains as one of the most widely used as the
pesticide. Solid-phase extraction (SPE) is an extraction method that uses solid and liquid
phase to isolate analyte from solution. It is a rapid and selective sample preparation. The
purpose of using the SPE is to purify, trace enrichment, solvent exchange and derivatization.
The advantages of the SPE are high recovery of analyte, concentration of analyte, ease of
automation and reduction of organic solvent consumption. The general procedure of SPE is
the loading of solution through SPE phase, wash away undesired components, and lastly
wash off the desired analytes with other solvent into the collection tube. Steps of SPE are
from column solvation and equilibration, sample loading, interference elution and lastly, the
analyte elution. The objectives of this experiment are to calculate the amount of chlorpyrifos
and each waste water sample and also the percentage of recovery.
PROCEDURE:

a) Filter Water Sample Through A Glass Fibre Filter Paper

b) Solid-phase Extraction Procedure

(i). C18 SPE cartridge was conditioned by passing 10 mL of methanol.

(ii). The cartridge was rinsed by passing 6 mL of deionized water without applying vacuum.
(iii). Then, the filtered water sample was passed through the preconditioned column using a
vacuum manifold at ~6 mL per min (about 48 drops min-1). The column should not be
allowed to dry during this sample enrichment step.

(iv). The column was dried by vacuum for 15 minutes.

(v). The interference was removed by eluting the column with 10 mL of deionized water and
the cartridge was dried by vacuum for 30 minutes. The cartridge was placed into tube rack.

(vi). After that, the pesticide was eluted using 5 mL of hexane. The sample was concentrated
to 1 mL by blowing down using gentle nitrogen and the sample were sealed by using parafilm
for GC analysis.

c) Instrument Set-up

Injector Temperature: 280oC

Detector Temperature: 300oC

Carrier Gas Flow Rate: 20.0 mL min-1 (Nitrogen)

Column Temperature: Initial Temperature 165oC for 3 minutes, increase to 260oC at 3oC
min-1 with final time of 2 minutes
d) Quantitative Analysis Of Chlorpyrifos

(i). 1 µL of sample was injected onto the column. The injection was repeated to get
reproducible peak areas.

(ii). Then, 1 µL of standard chlorpyrifos were injected and repeated to get reproducible peak
area.

(iii). The concentrations of chlorpyrifos in the sample were calculated using the data from
standard solution.

CALCULATION

1 mg/mL = 1000 ppm

X mg/mL = 100 ppm

 100+1
x=
1000

x = 0.0001 g/mL

MV mass
No. mole = =
1000 molar mass

MV
Mass chloropyrifos = × molar mass
1000

g
= ( 0.0001
mL )
(100 mL)
× 350.59 g/mol
1000

= 0.003506 g @ 0.3506 mg

Calculation of response factor standard:

amount
RF =
peak area

30
=
3.07805e4

= 9.7464e¯⁴ ppm/Hz*s

Amount of chlorpyrifos in sample (ppm):

Amount of chlorpyrifos = RF standard x peak area sample

= 9.7464e¯⁴ ppm/Hz*s × 1.20926e6 Hz*s

= 11.79 ppm
Percentage of recovery of sample:

amount of chlorpyrifos ∈sample

% recovery = × 100%
amount of chlorpyrifos∈standard

11.79 ppm
= × 100 %
30 ppm

= 39.3%

DISCUSSION

In this experiment, CG with Electron Capture detector is used because the analyte to be
analysed is halogenated compound. ECD only can detect analytes which contain
electronegative functional groups that can capture electrons as halogens, peroxides, quinones
and nitro groups. The disadvantage of ECD is it involve radioactive component. The amount
of chlorpyrifos in samples is calculated by using response factor calculation that base on the
standard compound. The amount of chlorpyrifos in each sample does not show big different,
it show that same procedure was carried out each sample. The retention time of chlorpyrifos
in standard is 5.516min while retention time of chlorpyrifos in sample is 5.602 min, the
retention time in sample we get from the retention time of standard in range. Then we
calculated of response factor for standard and we get 9.7464e¯⁴ ppm/Hz*s. We multiply
peak area of sample with respon factor to get the amount of chloropyrifos in sample to
calculated the average amount of chlorpyrifos. The average of chlorprifos in the sample is
5.895ppm, that is about half from the amount of chlorpyrifos in the standard. The percentage
recovery calculated 39.3% in average. In order to get higher amount of chlorpyrifos or the
percentage recovery, the SPE must be carried out carefully so it will extract more
chlorpyrifos efficiciently.

CONCLUSION
The average amount of chlorpyrifos in sample is 5.895ppm and the percentage recovery is
39.3%.

REFERENCE:

1. Nor’ashikin S., Ruziyati T., Mardiana S. (2012), Analytical Separation Methods

Laboratory Guide (2nd edition).

2. Azhar A., Dec (2014), Retrieved from

https://www.scribd.com/doc/249342246/ANALYSIS-OF-CHLORPYRIFOS-IN-WATER-
BY-SOLID-PHASE-EXTRACTION-SPE-AND-GAS-CHROMATOGRAPHY-
ELECTRON-CAPTURE-DETECTOR-GC-ECD

3. Faizzarul and Fadli M. Retrieved from

http://fareastzareul1994.blogspot.com/2018/05/experiment-5-analysis-of-chlorpyrifos.html?
m=1